使用磁共振成像实验性脑疟疾模型中的水肿发育和微血管病理学的体内追踪
Summary
We describe a mouse model of experimental cerebral malaria and show how inflammatory and microvascular pathology can be tracked in vivo using magnetic resonance imaging.
Abstract
Cerebral malaria is a sign of severe malarial disease and is often a harbinger of death. While aggressive management can be life-saving, the detection of cerebral malaria can be difficult. We present an experimental mouse model of cerebral malaria that shares multiple features of the human disease, including edema and microvascular pathology. Using magnetic resonance imaging (MRI), we can detect and track the blood-brain barrier disruption, edema development, and subsequent brain swelling. We describe multiple MRI techniques that can visualize these pertinent pathological changes. Thus, we show that MRI represents a valuable tool to visualize and track pathological changes, such as edema, brain swelling, and microvascular pathology, in vivo.
Introduction
疟疾是一个重大的全球健康问题。 1严重疟疾的特征部分是脑受累，往往是不良预后因素。疟疾传播高的地区，五岁以下儿童的脑梗死是常见的，并且是该年龄组疟疾相关死亡的主要原因。 1虽然积极的治疗可以挽救生命，但是特别是早期阶段的脑疟疾检测可能很困难。涉及脑疟疾的病理过程包括微血管破裂和脑水肿，可导致严重的脑肿胀。在本文中，我们提出了磁共振成像（MRI）协议，允许全脑脑实验性脑疟疾（ECM） 体内成像。全脑高分辨率成像方法在这种疾病中已经广泛利用不足，尽管对ECM如何在中枢起搏方面知之甚少神经系统或什么具体机制导致疾病。涵盖全脑的体内 MRI代表了一种重要的研究工具，可以更好地了解ECM病理学。 MRI能够评估全球大脑脑肿胀，最近已经被认为是ECM的重要预测因子，也是人脑疟疾的重要预测因素。 2，3严重的脑肿胀发生在致命疾病中，代表了ECM模型与人类疾病之间的几种病理特征之一，其特征是炎症和微血管改变。 4
可以通过感染致死的疟原虫疟原虫ANKA在CBA或C57BL小鼠中诱导ECM 。 ECM的发病通常发生在感染后6天和10天之间，并且导致拟合，共济失调，呼吸窘迫和昏迷，这导致说唱id死亡。 4快速鼠类昏迷和行为量表（RMCBS）是评估ECM临床症状的有益成绩。它由10个参数组成，每个评分从0到2，最大可能得分为20. 6最近，我们在ECM小鼠中RMCBS评分的严重程度和MRI证实的病理变化之间表现出良好的一致性。 7在本协议中，我们描述了小鼠的ECM诱导和ECM小鼠的体内磁共振成像。
Protocol
Representative Results
Discussion
在本文中，我们描述了一个全脑MRI方案来描绘实验性脑疟疾的变化。我们认为迄今为止，疟疾研究中MRI尚未得到充分利用，希望我们的方案能够帮助其他调查人员。我们想描述一些可能有帮助的附加要点。
如果严重恶心的小鼠成像，定位至关重要。由于颅内压升高，小鼠易死亡，因此颈椎不应伸展。麻醉也应保持在最低限度。如果轻度疾病的小鼠成像，MRI的最佳时间点是在?…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
衷心感谢Miriam Reinig的专家技术援助。 AH由海德堡大学医学院的博士后补助金资助。 MP由Else-Kröner-Fresenius基金会的纪念津贴支持。 AKM是德国感染研究中心（DZIF）DZIF学院的产假津贴的获得者。 JP是海德堡分子医学研究中心（HRCMM）职业发展奖学金的获得者。我们进一步感谢Julia M. Sattler和Friedrich Frischknecht提供了子孢子运动的典型电影。
Materials
| Isoflurane | Baxter | 1001747 | for anesthesia |
| Dotarem | Guebert | 1086923 | Gd-DTPA contrast agent; 0.5mmol/ml |
| Amira (Image Processing Program) | FEI Group | Version Amira 5.3.2 | |
| MATLAB | The MathWorks, Inc., | Release 2012b | |
| FDT toolbox | FMRIB's Software Library | http://www.fmrib.ox.ac.uk/fsl/fdt/index.html | |
Riferimenti
- World Health Organization. . World Malaria Report. , (2014).
- Seydel, K. B., et al. Brain swelling and death in children with cerebral malaria. N Engl J Med. 372 (12), 1126-1137 (2015).
- Penet, M. F., et al. Imaging experimental cerebral malaria in vivo: significant role of ischemic brain edema. J Neurosci. 25 (32), 7352-7358 (2005).
- de Souza, J. B., Riley, E. M. Cerebral malaria: the contribution of studies in animal models to our understanding of immunopathogenesis. Microbes Infect. 4 (3), 291-300 (2002).
- Curfs, J. H., van der Meide, P. H., Billiau, A., Meuwissen, J. H., Eling, W. M. Plasmodium berghei: recombinant interferon-gamma and the development of parasitemia and cerebral lesions in malaria-infected mice. Exp Parasitol. 77 (2), 212-223 (1993).
- Carroll, R. W., et al. A rapid murine coma and behavior scale for quantitative assessment of murine cerebral malaria. PLoS One. 5 (10), (2010).
- Hoffmann, A., et al. Experimental Cerebral Malaria Spreads along the Rostral Migratory Stream. PLoS Pathog. 12 (3), e1005470 (2016).
- Mueller, A. K., Behrends, J., Blank, J., Schaible, U. E., Schneider, B. E. An experimental model to study tuberculosis-malaria coinfection upon natural transmission of Mycobacterium tuberculosis and Plasmodium berghei. J Vis Exp. (84), e50829 (2014).
- Hynynen, K., McDannold, N., Sheikov, N. A., Jolesz, F. A., Vykhodtseva, N. Local and reversible blood-brain barrier disruption by noninvasive focused ultrasound at frequencies suitable for trans-skull sonications. Neuroimage. 24 (1), 12-20 (2005).
- Nag, N., Mellott, T. J., Berger-Sweeney, J. E. Effects of postnatal dietary choline supplementation on motor regional brain volume and growth factor expression in a mouse model of Rett syndrome. Brain Res. 1237, 101-109 (2008).
- Giri, S., et al. T2 quantification for improved detection of myocardial edema. J Cardiovasc Magn Reson. 11, 56 (2009).
- Engwerda, C., Belnoue, E., Gruner, A. C., Renia, L. Experimental models of cerebral malaria. Curr Top Microbiol Immunol. 297, 103-143 (2005).
- Zhao, H., et al. Olfactory plays a key role in spatiotemporal pathogenesis of cerebral malaria. Cell Host Microbe. 15 (5), 551-563 (2014).
- Nacer, A., et al. Experimental cerebral malaria pathogenesis–hemodynamics at the blood brain barrier. PLoS Pathog. 10 (12), e1004528 (2014).
- Nacer, A., et al. Neuroimmunological blood brain barrier opening in experimental cerebral malaria. PLoS Pathog. 8 (10), e1002982 (2012).
- Pai, S., et al. Real-time imaging reveals the dynamics of leukocyte behaviour during experimental cerebral malaria pathogenesis. PLoS Pathog. 10 (7), e1004236 (2014).
- Shaw, T. N., et al. Perivascular Arrest of CD8+ T Cells Is a Signature of Experimental Cerebral Malaria. PLoS Pathog. 11 (11), e1005210 (2015).
- Potchen, M. J., et al. Acute brain MRI findings in 120 Malawian children with cerebral malaria: new insights into an ancient disease. AJNR Am J Neuroradiol. 33 (9), 1740-1746 (2012).
